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Research on genetic diversity of Chinese population of the crested ibis.

中国朱鹮种群遗传多样性研究.

基本信息

批准号：
13575003
负责人：
TAMATE Hidetoshi
金额：
$ 6.34万
依托单位：
Yamagata University (2003)Ishinomaki Senshu University (2001-2002)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13575003/
关键词：
China endangered species genetic diversity mitochondrial DNA molecular phylogeny conservation population genetics PCR AFLP 野生動物遺伝子分析

项目摘要

The objective of this study is to measure the level of genetic diversity of a population of the crested ibis, an endangered species, in China, and to establish genetic markers that can be utilized for monitoring inbreeding status of individual animals. For this purpose, we collected DNA sapmles from dead animals and examine genetic variations among the samples. First, we determined a partial nucleotide sequence of mitochondrial DNA (mtDNA), and analyzed phylogenetic relationship between the crested ibis and its related species, Our data shows that the crested ibis is most closely related to Plegadis falcinellus, Glossy Ibis. We, however, found no variation in mtDNA sequence among individuals. On the other hand, variations at individual level were found by the method of amplified fragment length polymorphism (AFLP). In AFLP experiment, we used EcoRI and MseI restriction endonucleases to generate DNA fragments. Then we used EcoRI-restricted primer and MseI-restricted primer to amplify selected DNA fragments At final step of AFLP, we used 12sets of selective primers to limit the number of DNA bands in the analysis. Two sets of the primers gave different fragment pattern for individual samples, The result suggest that AFLP would be applicable to determine pedigree structures in the crested ibis population if more AFLIP markers are identified. Other nuclear markers such as microsatellite and non-coding sequences were also identified in the sample population, however, no variation was found among samples so far. We also sequenced a part of major histocompatibility complex loci, but failed to find genetic variations. Those data suggest that the population of the Chinese crested ibis is tinder extreme inbreeding condition and the level of genetic diversity is greatly reduced, although some variations still exist.

本研究的目的是测定中国濒危物种朱鹮种群的遗传多样性水平，并建立可用于监测个体近交状况的遗传标记。为此，我们从死亡动物中收集DNA样本，并检查样本中的遗传变异。首先测定了朱鹮线粒体DNA（mtDNA）的部分核苷酸序列，并分析了朱鹮与其近缘种的系统发育关系。然而，我们没有发现个体之间的mtDNA序列的变化。另一方面，通过扩增片段长度多态性（AFLP）方法，发现了个体水平上的变异。在AFLP实验中，我们使用EcoRI和MseI限制性内切酶来产生DNA片段。然后用EcoRI限制性引物和MseI限制性引物对筛选出的DNA片段进行扩增，最后用12对选择性引物限制扩增带数。结果表明，如果能筛选出更多的AFLP标记，AFLP技术可用于朱鹮群体的系谱结构分析。其他核标记如微卫星和非编码序列也在样本群体中被发现，但迄今为止，没有发现样本之间的变异。我们还对部分主要组织相容性复合体基因座进行了测序，但未能发现遗传变异。这些数据表明，中国朱鹮种群处于极端近交状态，遗传多样性水平大大降低，但仍存在一定的变异。